植物乳杆菌AKU 1009a中催化多不饱和脂肪酸饱和代谢第一步的亚油酸Δ9水合酶的特性分析。
Characterization of the linoleic acid Δ9 hydratase catalyzing the first step of polyunsaturated fatty acid saturation metabolism in Lactobacillus plantarum AKU 1009a.
作者信息
Takeuchi Michiki, Kishino Shigenobu, Hirata Akiko, Park Si-Bum, Kitamura Nahoko, Ogawa Jun
机构信息
Division of Applied Life Sciences, Graduate School of Agriculture, Kyoto University, Kitashirakawa-oiwakecho, Sakyo-ku, Kyoto 606-8502, Japan.
Division of Applied Life Sciences, Graduate School of Agriculture, Kyoto University, Kitashirakawa-oiwakecho, Sakyo-ku, Kyoto 606-8502, Japan; Laboratory of Industrial Microbiology, Graduate School of Agriculture, Kyoto University, Kitashirakawa-oiwakecho, Sakyo-ku, Kyoto 606-8502, Japan.
出版信息
J Biosci Bioeng. 2015 Jun;119(6):636-41. doi: 10.1016/j.jbiosc.2014.10.022. Epub 2014 Dec 1.
Linoleic acid Δ9 hydratase, which is involved in linoleic acid saturation metabolism of Lactobacillus plantarum AKU 1009a, was cloned, expressed as a his-tagged recombinant enzyme, purified with an affinity column, and characterized. The enzyme required FAD as a cofactor and its activity was enhanced by NADH. The maximal activities for the hydration of linoleic acid and for the dehydration of 10-hydroxy-cis-12-octadecenoic acid (HYA) were observed at 37 °C in buffer at pH 5.5 containing 0.5 M NaCl. Free C16 and C18 fatty acids with cis-9 double bonds and 10-hydroxy fatty acids served as substrates for the hydration and dehydration reactions, respectively. The apparent Km value for linoleic acid was estimated to be 92 μM, with a kcat of 2.6∙10(-2) s(-1) and a Hill factor of 3.3. The apparent Km value for HYA was estimated to be 98 μM, with a kcat of 1.2∙10(-3) s(-1).
参与植物乳杆菌AKU 1009a亚油酸饱和代谢的亚油酸Δ9水合酶被克隆，表达为带组氨酸标签的重组酶，用亲和柱纯化并进行了表征。该酶需要FAD作为辅因子，其活性被NADH增强。在含0.5 M NaCl、pH 5.5的缓冲液中，于37 °C观察到亚油酸水合和10-羟基-顺式-12-十八碳烯酸（HYA）脱水的最大活性。带有顺式-9双键的游离C16和C18脂肪酸以及10-羟基脂肪酸分别作为水合和脱水反应的底物。亚油酸的表观Km值估计为92 μM，kcat为2.6∙10(-2) s(-1)，希尔系数为3.3。HYA的表观Km值估计为98 μM，kcat为1.2∙10(-3) s(-1)。
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